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Cell in situ zymography: an in vitro cytotechnology for localization of enzyme activity in cell culture

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Abstract

In situ zymography is a unique technique for detection and localization of enzyme–substrate interactions majorly in histological sections. Substrate with quenched fluorogenic molecule is incorporated in gel over which tissue sections are mounted and then incubated in buffer. The enzymatic activity is observed in the form of fluorescent signal. With the advancements in the field of biological research, use of in vitro cell culture has become very popular and holds great significance in multiple fields including inflammation, cancer, stem cell biology and the still emerging 3-D cell cultures. The information on analysis of enzymatic activity in cell lines is inadequate presently. We propose a single-step methodology that is simple, sensitive, cost-effective, and functional to perform and study the ‘in position’ activity of enzyme on substrate for in vitro cell cultures. Quantification of enzymatic activity to carry out comparative studies on cells has also been illustrated. This technique can be applied to a variety of enzyme classes including proteases, amylases, xylanases, and cellulases in cell cultures.

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Acknowledgments

This work was supported by the research grant awarded to Dr. Vibha Rani by the Department of Science and Technology, Government of India (SR/FT/LS-006/2009: Sept 4, 2009). We also acknowledge Jaypee Institute of Information Technology, Deemed to be University, for the infrastructural support.

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Authors and Affiliations

  1. Department of Biotechnology, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, 201307, Uttar Pradesh, India

    Aastha Chhabra, Astha Jaiswal, Umang Malhotra, Shrey Kohli & Vibha Rani

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  1. Aastha Chhabra
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  2. Astha Jaiswal
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  3. Umang Malhotra
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  4. Shrey Kohli
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  5. Vibha Rani
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Correspondence to Vibha Rani.

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Editor: T. Okamoto

The authors Aastha Chhabra, Astha Jaiswal, Umang Malhotra, Shrey Kohli, and Vibha Rani have contributed equally to this paper.

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Chhabra, A., Jaiswal, A., Malhotra, U. et al. Cell in situ zymography: an in vitro cytotechnology for localization of enzyme activity in cell culture. In Vitro Cell.Dev.Biol.-Animal 48, 463–468 (2012). https://doi.org/10.1007/s11626-012-9529-5

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  • DOI: https://doi.org/10.1007/s11626-012-9529-5

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